IL-2基因修饰人骨肉瘤细胞的制备及骨肉瘤患者IL-2和sIL-2R测定

The Preparation of Human Osteosarcoma Cells Modified by IL- 2 Gene and the Determination of IL -2 and sIL -2R in Patients with Osteosarcoma

目的ＩＬ－２基因修饰人骨肉瘤细胞的制备及骨肉瘤患者ＩＬ－２、ｓＩＬ－２Ｒ水平测定。方法用逆转录病毒载体将人ＩＬ－２基因插入人骨肉瘤细胞系制备成ＩＬ－２基因修饰骨肉瘤细胞；用放射免疫法及ＥＬＩＳＡ法检测骨肉瘤患者血清内ＩＬ－２、ｓＩＬ－２Ｒ水平。结果经Ｇ４１８筛选和ＰＣＲ鉴定后制备成功人骨肉瘤ＩＬ－２基因修饰细胞，并证实其有分泌ＩＬ－２功能，用ＭＴＴ法测得其培养上清中ＩＬ－２表达量为每１ｘ１０５细胞２４小时５０－８００ｕ。骨肉瘤患者免疫功能调查发现其血清ＩＬ－２水平明显低于正常，而ｓＩＬ－２Ｒ则明显高于正常，术后两者可逐渐正常。结论 制备成功ＩＬ－２基因修饰骨肉瘤细胞并发现骨肉瘤患者ＩＬ－２水平低下，为进行骨肉瘤基因治疗创造条件。

Objective To produce human osteosarcoma cells modified by IL - 2 gene and investigatethe level of IL - 2 and sIL - 2R in patients with osteosarcoma. Methods The human IL- 2 gene was in-serted into human osteosarcoma cells with retroviral vector and the tumor vaccine was produced. IL-2 andsIL -2R level in patients with osteosarcoma was determined with radioimmunoassay and ELISA. ResultsThe human osteosarcoma cells modified by IL - 2 gene was produced successfully after G418 resistence se-lection and analysed by PCR; they could secrete IL - 2 in vitro. The production of IL - 2 was 50 ￣ 800 u per1 × 10 cells every 24 h. The data about patients' immune state indicated that IL - 2 was lower than normalcontrol and sIL - 2R higher. Conclusion The success in production of the human osteosarcoma cells mod-ified by IL - 2 gene makes it possisle to carry out further reseach in gene therapy for human osteosarcoma.